Structure and characterization of an extracellular polysaccharide from Paenibacillus polymyxa 88A.

Peptides from scorpion venom have been previously studied for use in the prevention and treatment of various types of cancer in folk medicine. The present study investigated the anti-proliferative effects and mechanisms of the low molecular weight (~3 kDa) BmK scorpion venom peptides (LMWSVP) on human hepatoma (SMMC 7721) and cervical carcinoma (HeLa) cells. The data indicated that LMWSVP inhibited the growth of SMMC 7721 cells, but had no effect on the growth of HeLa cells. SMMC 7721 cells were more sensitive, with a higher affinity, to LMWSVP as compared with HeLa cells. In addition, LMWSVP induced apoptosis of SMMC 7721 cells by upregulating the expression of caspase-3 and downregulating the expression of Bcl-2. These data provide an experimental basis for further purification and application of LMWSVP for use as an anti-tumor drug in clinical trials.

In this study, a novel taxol-producing endophytic fungus, strain F3, was isolated from the fruits of Taxus cuspidata and identified as Alternaria alternata according to its macroscopic and microscopic traits and sequence analysis of internal transcribed spacer (ITS). The presence of taxol was detected by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) and confirmed by ultra-high-performance liquid chromatography-electrospray coupled to tandem mass spectrometry (UPLC-ESI-MS/MS) and nuclear magnetic resonance (NMR). The fermentation parameters of strain F3 were then optimized for high taxol production. The maximum taxol yield of 195.4µg L-1 by A. alternata F3 was observed in 200-mL yeast peptone dextrose (YPD) broth, at an initial pH value of 6.0, supplemented with 0.1g L-1 sodium acetate, 0.25g L-1 salicylic acid, and 0.00125g L-1 silver nitrate and inoculum size 2%, and incubated at 28°C and 150rpm for 8days, which was 2.12-fold compared with the initial yield of taxol. Also, fungal taxol exhibited antitumor activity towards human lung carcinoma (A549) cell line and human cervical carcinoma (Hela) cell line with IC50 values of 3.98µgmL-1 and 0.35µgmL-1. Overall, this is the first report on taxol-producing endophytic fungus isolated from the fruits of Taxus. This study offers a novel source for the production of taxol for anticancer treatment.

Histone demethylase JHDM1D (also known as KDM7A) modifies the level of methylation in histone and participates in epigenetic gene regulation; however, the role of JHDM1D in tumor progression is unknown. Here, we show that JHDM1D plays a tumor-suppressive role by regulating angiogenesis. Expression of JHDM1D was increased in mouse and human cancer cells under long-term nutrient starvation in vitro. Expression of JHDM1D mRNA was increased within avascular tumor tissue at the preangiogenic switch, along with increased expression of angiogenesis-regulating genes such as Vegf-A. Stable expression of JHDM1D cDNA or siRNA silencing of JHDM1D in cancer cells did not affect cell proliferation, anchorage-independent cell growth, or cell cycle progression in vitro. Notably, JHDM1D-expressing mouse melanoma (B16) and human cervical carcinoma (HeLa) cells exhibited significantly slower tumor growth in vivo compared with the original cells. This reduction in tumor growth was associated with decreased formation of CD31(+) blood vessels and reduced infiltration of CD11b(+) macrophage linage cells into tumor tissues. Expression of multiple angiogenic factors such as VEGF-B and angiopoietins was decreased in tumor xenografts of JHDM1D-expressing B16 and HeLa cells. Our results provide evidence that increased JHDM1D expression suppressed tumor growth by down-regulating angiogenesis under nutrient starvation.

Human growth hormone gene expression in rat but not human non-pituitary cells after stable gene transfer

ADP-Ribosylation of Heterogeneous Ribonucleoproteins in HeLa Cells

A water-soluble zinc(II) phthalocyanine (ZnPc1) substituted with 1-(3-methyl) imidazoliumylethyloxy on the periphery was synthesized, and its subcellular localization and photodynamic activities were studied. ZnPc1 exhibited an almost exclusive mitochondrial-localizing property in human cervical carcinoma (HeLa) cells. Remarkable photocytotoxicity but low dark cytotoxic properties were observed for ZnPc1 in four different cancer cell lines. Photodynamic treatment (PDT) with ZnPc1 resulted in the generation of reactive oxygen species, a collapse of the mitochondrial membrane potential and chromatin condensation, all important hallmarks of apoptosis. The results are consistent with its mitochondrial localization property and qualify ZnPc1 as a promising PDT agent.

We have synthesized ferrocene-appended Y-shaped methoxyphenyl-substituted imidazole derivatives 2-ferrocenyl-4,5-bis(4-methoxyphenyl)-1H-imidazole (1) and 2-ferrocenyl-4,5-bis((E)-4-methoxystyryl)-1H-imidazole (2), and characterized them using analytical and spectroscopic techniques (1H, 13C NMR, FT-IR, and HR-Mass). Optical properties, including absorption, emission, quantum yield, and aggregation-induced emission (AIE), were demonstrated. The emission, ranging from weak to enhanced, was achieved through the AIE process in CH3CN/H2O mixtures by restricting intramolecular rotation (RIR). In the aggregated state, the quantum efficiency of chromophores 1 and 2 increases threefold and twofold, respectively, compared to the pure CH3CN solution. Utilizing the AIE state, these chromophores were further employed to detect hydrogen peroxide (H2O2) in a mixed aqueous medium via a fluorescence turn-on/off approach. This was conducted with water fractions (fw) of 80 % for chromophore 1 and 70 % for chromophore 2, revealing a highly specific and sensitive fluorescence quenching response to H2O2 in the aqueous mixture. The response is rapid within a linear range of 0–110 µM and 0–80 µM, with detection thresholds of 24.7 nM for chromophore 1 and 30.9 nM for chromophore 2, respectively. The limited detection capability is attributed to the absence of an alkene group in the chromophore, leading to low sensitivity due to the chromophore's high molecular rigidity, which impedes molecular motion. Moreover, chromophores 1 and 2 are non-cytotoxic (cell viability above 80 %), enabling them to detect intracellular H2O2 in human cervical carcinoma (HeLa) cells through fluorescence bio-imaging. Notably, the significant bright green fluorescence is quenched upon the addition of H2O2. Additionally, DFT/B3LYP calculations were used to explore the HOMO and LUMO energy levels and charge distribution of the optimized structure. The calculated HOMO and LUMO energies were compared with the experimentally obtained redox potential values.

Chronic toxicity and cytotoxicity of synthetic pyrethroid insecticide cis-bifenthrin

Phosphorylation of heat shock protein 27 antagonizes TNF-α induced HeLa cell apoptosis via regulating TAK1 ubiquitination and activation of p38 and ERK signaling

The purpose of this study was to develop nanoparticles made of cholesterol-conjugatedcarboxymethyl curdlan (CCMC) entrapping epirubicin (EPB) and establish their invitro and in vivo potential. CCMC was synthesized and characterized by Fouriertransform infrared spectra (FT-IR) and proton nuclear magnetic resonance spectra (1H NMR). The degrees of substitution (DS) of the cholesterol moiety were 2.3, 3.5 and 6.4,respectively. EPB-loaded CCMC-3.5 nanoparticles were prepared by the remote loadingmethod. The physicochemical characteristics, drug loading efficiency and drug releasekinetics of EPB-loaded CCMC-3.5 nanoparticles were characterized. The in vitrorelease profiles revealed that EPB release was sensitive to the pH as well as thedrug loading contents. The cellular cytotoxicity and cellular uptake were accessedby using human cervical carcinoma (HeLa) cells. The EPB-loaded CCMC-3.5nanoparticles were found to be more cytotoxic and have a broader distribution within thecells than the free EPB. The in vivo pharmacokinetics and biodistribution wereinvestigated after intravenous injection in rats. Promisingly, a 4.0-fold increase inthe mean residence time (MRT), a 4.31-fold increase in the half-life time and a6.69-fold increase in the area under the curve of EPB were achieved for the EPB-loaded CCMC-3.5 self-assembled nanoparticlescompared with the free EPB. The drug level was significantly increased in liver at 24 and72 h; however, it decreased in heart at 8 and 24 h compared with the free EPB. The in vivoanti-tumor study indicated that the EPB-loaded CCMC-3.5 self-assembled nanoparticlesshowed greater anti-tumor efficacy than the free EPB. Taken together, the novelCCMC self-assembled nanoparticles might have potential application as anti-cancerdrug carriers in a drug delivery system due to good results in vitro and in vivo.

Two non-polar fractions viz. hexane (Hex-LI) and chloroform fraction (CHCl3-LI) of Lawsonia inermis were studied for their antiproliferative potential in various cancer cell lines viz. HeLa, MCF-7, A549 and C6 glioma cells. Both the fractions showed more than 60% of growth inhibition in all the tested cell lines at highest tested concentration. In clonogenic assay, different concentrations of Hex-LI and CHCl3-LI decreased the number and size of colonies as compared to control in HeLa cells. The apoptotic effects as nuclear condensation, fragmentation were visualized with Hoechst-33342 staining of HeLa cells using confocal microscope. Both fractions induced apoptotic cell death in human cervical carcinoma (HeLa) cells as evident from flow cytometric analysis carried out using Annexin V-FITC and propidium iodide dyes. CHCl3-LI treated cells significantly induced apoptosis (25.43%) in comparison to control. Results from Neutral Comet assay demonstrated that both fractions induced double stranded breaks (DSB's) in HeLa cells. Our data indicated that Hex-LI and CHCl3-LI treated cells showed significant increase of 32.2 and 18.56% reactive oxygen species (ROS) levels in DCFH-DA assayrespectively. Further, experimental studies to decipher exact pathway via which these fractions induce cell death are in progress.

Anticancer activity of grape and grape skin extracts alone and combined with green tea infusions

BackgroundCervical cancer is one of the most common gynecological malignancies. Previous studies have shown that the ethanol extract of Sophora moorcroftiana seeds (EESMS) possesses an antiproliferative effect on several tumors in vitro. Therefore, in this study, we assessed the impact of EESMS on human cervical carcinoma (HeLa) cell proliferation.MethodsThe proliferation and apoptotic effects of HeLa cells treated with EESMS were evaluated using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay, dual acridine orange/ethidium bromide double staining, flow cytometry, and western blotting. Single-cell level atomic force microscopy (AFM) was conducted to detect the mechanical properties of HeLa cells, and proteomics and bioinformatics methods were used to elucidate the molecular mechanisms of EESMS.ResultsEESMS treatment inhibited HeLa cell proliferation by blocking the G0/G1 phase, increasing the expression of Caspase-3 and affecting its mechanical properties, and the EESMS indicated no significant inhibitory effect on mouse fibroblasts L929 cell line. In total, 218 differentially expressed proteins were identified using two-dimensional electrophoresis, and eight differentially expressed proteins were successfully identified using matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry. The differentially expressed proteins were involved in various cellular and biological processes.ConclusionThis study provides a perspective on how cells change through biomechanics and a further theoretical foundation for the future application of Sophora moorcroftiana as a novel low-toxicity chemotherapy medication for treating human cervical cancer.

We investigated the coordinative behaviour of the phosphanylthiosemicarbazone ligand 2‐[2‐(diphenylphosphanyl)benzylidene]‐N‐phenylthiosemicarbazone (HLPPh) towards coinage MI metal ions (M = Cu, Ag, Au). The complexes obtained, with formulae [Cu(HLPPh)2]Br (1), [Cu(HLPPh)2]I·H2O (2), [Ag(HLPPh)(NO3)]·3H2O (3) and [Au2(HLPPh)2]·3CH3OH (4), were satisfactorily characterized by elemental analysis, IR and 1H/31P NMR spectroscopy as well as ESI mass spectrometry. In addition, we obtained the crystal structure of the complex [Au2(μ‐S,P‐HLPPh)2]Cl2 (5), which is the first example of a homoleptic thiosemicarbazone AuI complex, because the AuI ion has been stabilized by exclusively using a phosphanylthiosemicarbazone ligand. We also performed preliminary luminescence studies for these complexes, as well as the screening of its biological activity against human cervical carcinoma (HeLa) cells. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2008)

A series of heterocyclic compounds containing spiro-fused pyrrolo[3,4-a]pyrrolizine and tryptanthrin framework have been synthesized and studied as potential antitumor agents. Antiproliferative activity of synthesized products was screened against human erythroleukemia (K562) and human cervical carcinoma (HeLa) cell lines. Spiroadducts with all cis bridge-protons of pyrrolo[3,4-a]pyrrolizine moiety were more active in all cases. Replacement of hydrogen atom of pyrrole moiety by either alkyl or aryl group leads to significant decreasement in activity of both formed cycloadducts. In agreement with the DNA cytometry studies, the tested compounds have achieved significant cell-cycle perturbation with higher accumulation of cells in G2/M phase and induce apoptosis. Using confocal microscopy, we found that with synthesized products treatment of HeLa cells, actin filaments disappeared, and granular actin was distributed diffusely in the cytoplasm in 76-91% of cells. We discovered that HeLa cells after treatment with screened compounds significantly reduced the number of cells with filopodium-like membrane protrusions (from 63 % in control cells to 29% after treatment) and decrease in cell motility. All obtained data make it possible to assume that spiro-fused pyrrolo[3,4-a]pyrrolizine and tryptanthrin framework may be considered as a new promising pharmacophore unit for further screenings.